To determine the molecular profile of the multidrug-resistant (MDR) strains of Mycobacterium tuberculosis isolated in Senegal. From 2011 to 2014, 43 MDR strains isolated at the National Reference Laboratory for Mycobacteria of the National Antituberculosis Program (NAP) were characterized by spoligotyping. Nine different spoligotypes were identified, three of which were unique, the other six forming clusters of two to nine isolates. The three principal families identified were the T superfamily (12/43, 27.90%), Beijing (9/43, 20.93%), and Latino-American-Mediterranean (LAM; 3/43, 6.97%). Twelve isolates could not be attributed to a particular lineage. The strains causing multidrug-resistant tuberculosis in Senegal belong to three main families: the T superfamily, Beijing, and LAM.